4 years ago

Three-Dimensional Localization of an Individual Fluorescent Molecule with Angstrom Precision

Three-Dimensional Localization of an Individual Fluorescent Molecule with Angstrom Precision
Tsuyoshi Matsuda, Keisuke Wakao, Michio Matsushita, Satoru Fujiyoshi, Taku Furubayashi, Fuyuki Ishikawa, Mahito Sadaie, Takamitsu Hosoya, Isao Kii, Kazuya Motohashi, Nobuhiro Hayashi
Among imaging techniques, fluorescence microscopy is a unique method to noninvasively image individual molecules in whole cells. If the three-dimensional spatial precision is improved to the angstrom level, various molecular arrangements in the cell can be visualized on an individual basis. We have developed a cryogenic reflecting microscope with a numerical aperture of 0.99 and an imaging stability of 0.05 nm in standard deviation at a temperature of 1.8 K. The key optics to realize the cryogenic performances is the reflecting objective developed by our laboratory. With this cryogenic microscope, an individual fluorescent molecule (ATTO647N) at 1.8 K was localized with standard errors of 0.53 nm (x), 0.31 nm (y), and 0.90 nm (z) when 106 fluorescence photons from the molecule were accumulated in 5 min.

Publisher URL: http://dx.doi.org/10.1021/jacs.7b03899

DOI: 10.1021/jacs.7b03899

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